猪非结核分枝杆菌的分离与鉴定

采集结核菌素变态反应阴性猪的颌下淋巴结共计550份,肠系膜淋巴结共计550份,运用实验室常规检查、分离培养基分离培养、生长特性试验、鉴别培养基试验、产色试验、生物化学方法和分子生物学方法,对集约化养殖的猪群中非结核分枝杆菌（NTM）开展分离与鉴定研究。最终得到阳性结果14份,阳性率1.27%,其中颌下淋巴结6份,阳性率1.09%,肠系膜淋巴结8份,阳性率1.45%。分离株鉴定结果：2株为胃分枝杆菌,3株为胞内分枝杆菌,3株为偶发分枝杆菌,2株为牛分枝杆菌,4株为浅黄分枝杆菌。根据所得数据可以得知,结核阴性猪中NTM的流行情况不容乐观,其存在必定会对猪养殖业造成负面影响,试验为NTM的预防和中国消除结核病计划提供了理论依据。     

牛非结核分枝杆菌分离与鉴定

本试验对从吉林省不同区域的7个城市采集的503份牛颌下淋巴结样品和497份牛肠系膜淋巴结样品进行非结核分枝杆菌的流行情况的调查,样品均采自结核(副结核)变态反应阴性牛只,调查中得到阳性样品25份,阳性率2.5%,其中颌下淋巴结阳性率1.59%,肠系膜淋巴结阳性率3.42%,成功分离扩增培养出14株非结核分枝杆菌,并进行了分类鉴别,得到母牛分枝杆菌1株,龟分枝杆菌脓肿亚种2株,苏加分枝杆菌2株,偶发分枝杆菌1株,蟾蜍分枝杆菌6株,胞内分枝杆菌2株.调查结果表明:非结核分枝杆菌单独感染的情况不容乐观,应加强对非结核分枝杆菌的检验检疫工作.     

结核变态反应阳性牛的非典型性分枝杆菌的分离与鉴定

本试验从６９头结核变态反应阳性牛中采取病料２０２份进行分枝杆菌，特别是非典型分枝杆菌的分离与鉴定，共获２６株分枝杆菌，除５株分枝杆菌外，其它全是非典型分枝杆菌。在这些非典型分杆杆菌中，５株偶发分枝杆菌，６株瘰疠分枝杆菌，５株鸟－胞内分枝杆菌，２株副结核分枝杆菌和１株ＲｕｎｙｏｎⅡ群。从病料看，有病变的分离率为４１．２％，无病变的为７．１％。在三种淋巴结病料中，以肠系膜淋巴结的分离率为最高，颌下淋巴     

荧光定量RT-PCR方法检测HP-PRRSV JXA1分离株在藏猪、藏梅猪和大约克夏猪体内的动态分布

为了探明高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)JXA1分离株感染藏猪、藏梅猪和大约克夏猪后,病毒在猪体内的分布规律,本研究利用荧光定量RT-PCR技术检测3个品种猪感染4×105TCID50/m L的JXA1分离株后0、4、7和14 d体内的15个器官(心、肝、脾、肺、肾、大脑、小肠、颌下腺、颌下淋巴结、甲状腺、胸腺、支气管淋巴结、肠系膜淋巴结、腹股沟淋巴结和扁桃体)的病毒载量。结果表明,在整个感染期间,藏梅猪和大约克夏猪的15个器官中均检测到JXA1分离株,而藏猪的心、肝、脾、肾、小肠、颌下腺和甲状腺以及感染后14 d的脑中未检测到JXA1分离株;在JXA1分离株感染3个品种猪4、7和14 d,藏猪病毒载量最多的器官分别是扁桃体、腹股沟淋巴结和颌下淋巴结,藏梅猪分别是脾、脾和肺,大约克夏猪分别为胸腺、肺和颌下淋巴结。本研究证实了JXA1分离株在3个品种猪体内的分布规律和器官嗜性均存在明显差异。     

猪的抗酸菌病

猪的抗酸菌病(Mycobacterial Infectiou)是由分枝杆菌属感染而发生的慢性疾病。主要经口感染,并在头颈部或肠系膜淋巴结形成局灶性脓肿或干酪样坏死。发生原因:本病的发生,因猪的饲养环境而异。一般在牛结核多发区饲养的猪分离出牛(型)结核分     

奶水牛牛分枝杆菌的分离与鉴定

从患疑似结核病的广西奶水牛群中进行牛分枝杆菌的分离和鉴定。共收集了238份临床样品(鼻黏液及牛奶),病料经1.25 mol/mL NaOH溶液预处理后,接种罗氏培养基进行细菌分离。分离菌经进一步纯化后进行抗酸性染色并镜检,镜检后判为阳性的细菌再接种到鉴别培养基上进行鉴别培养,同时,应用PCR方法对镜检为阳性的细菌进行进一步的鉴别检验。结果收集的所有临床样品,经37℃培养后共获得细菌12株。这12株菌经抗酸性染色后镜检都为分枝杆菌阳性,进一步经鉴别培养基鉴定12株菌中有2株为牛分枝杆菌,其余均为非典型分枝杆菌。2株牛分枝杆菌经PCR方法鉴定得以确诊。     

猪轮状病毒与肠杆菌、猪丹毒杆菌混合感染的诊断

为查明毕节市某规模化种猪场腹泻流行的原因,通过流行病学调查、病理剖检、RT-PCR检测、细菌分离培养和药物敏感性试验对发病的10头新生仔猪进行诊断。结果:病理变化为颌下淋巴结出血和肠系膜淋巴结充血;猪轮状病毒RTPCR检测为阳性;细菌分离鉴定为革兰氏阴性肠杆菌和革兰氏阳性猪丹毒杆菌,对穿心莲等药物较为敏感。结论:猪腹泻由猪轮状病毒、肠杆菌和猪丹毒杆菌混合感染所致。     

西藏部分地区布鲁氏菌的分离和菌型鉴定

从西藏浪卡子、贡嘎等县11个乡收集牛、羊流产胎儿176头(只),分离出布鲁氏菌17株,连同保存的8株,共25株,经种型鉴定。其中,羊种菌(1、2、3型)21株,占鉴定菌株的84％;猪1型菌和未定型的菌各2株,均占鉴定菌株的8％。进一步证明,布鲁氏菌在西藏是以羊种菌为主,主要宿主有牛、羊和猪。     

猪抗酸菌病

1904年Webef和Bofinger首次从猪分离出分枝杆菌。此后,世界各国先后从猪淋巴结的结核样病变中分离出多种抗酸性分枝杆菌。因它可引起猪慢性传染病,故称为猪抗酸菌病,也叫猪分枚杆菌病。近年来,据国外报道,由人、牛结核菌所引起的猪结核病在逐渐减少,而由非典型分枝杆菌所引起的猪抗酸菌病则明显地增加。     

禽分枝杆菌在几种培养基上生长状况的初步观察(摘要)

实验对甘肃农业大学传染病教研组保存的长达5～31个月的127株鸡结核病病原进行了继代培养,结果表明,4℃冰箱保存23～31个月,细菌均未生长,保存期在9个月内的部分菌株明显生长,对其中10株细菌初步鉴定,有5株可确认为禽分枝杆菌.禽分枝杆菌在12种培养基上的生长状况以Lowenstain—Jensten 氏培养基为佳;琼脂培养基     

PATHOGENICITY FOR CATTLE OF ATYPICAL MYCOBACTERIA ISOLATED FROM FERAL PIGS AND CATTLE AND THE CORRELATION OF LESIONS WITH TUBERCULIN SENSITIVITY

Two experiments involving the inoculation of cattle with atypical mycobacteria are described. In the first experiment groups of 5 cattle were inoculated either subcutaneously or into a mesenteric lymph node with a strain of M. scrofulaceum or M. intracellulare. Four weeks and 10 weeks after inoculation the cattle were tuberculin tested with bovine PPD, avian PPD and homologous PPDs. The pathological changes observed were similar within each group of cattle inoculated with the same strain of mycobacteria. A significant interaction was demonstrated between the strain and the route of inoculation. In the second experiment 17 cattle were similarly inoculated by either of the two routes with 1 of 6 strains of M. intracellulare, a strain of M. scrofulaceum or a strain of Runyon Group IV, all of which had been isolated from feral pigs, or a strain of M. intracellulare of bovine origin. Tuberculin tests were carried out after 4 weeks and 10 weeks. Only the isolate from a bovine lymph node produced a significant level of sensitivity to bovine PPD. Cultural isolation of the mycobacteria from autopsy material was not correlated with the presence of macroscopic lesions nor with sensitivity to bovine PPD. The response to bovine PPD of cattle infected with these atypical mycobacteria decreased between 48 h and 96 h after injection of the tuberculins. As the maximum difference in the response to bovine and avian tuberculins occurs at 72 h a comparative tuberculin test should be read at this time to eliminate non-specific reactors.     

RESPONSE OF CATTLE TO INOCULATION WITH ATYPICAL MYCOBACTERIA OF BOVINE ORIGIN

SUMMARY Ten strains of atypical mycobacteria originally isolated from cattle were inoculated into cattle. Each strain was injected subcutaneously into one animal and into a mesenteric lymph node of another. Four weeks and 10 weeks after inoculation the cattle were tuberculin tested with bovine PPD, avian PPD and the appropriate homologous PPD. Three strains produced a significant level of sensitivity to bovine PPD at the 4-week test but by the 10-week test no animal gave a significant response. The sensitivity to all tuberculins was less at the 10-week test than at the 4-week test. At both tests the response to avian PPD was equal to or exceeded that to bovine PPD. Of 4 strains originally from cattle sensitive to mammalian tuberculin only 2 produced sensitivity of bovine PPD in this experiment. Cultural isolation of mycobacteria from necropsy material was correlated neither with sensitivity to bovine PPD nor with the presence of lesions.     

猪胴体淋巴结中金黄色葡萄球菌的分离与鉴定

随机采取西宁市某生猪屠宰场猪胴体淋巴结60份,进行细菌常规分离培养,结果分离出金黄色葡萄球菌菌株40株,表明该批猪胴体中的金黄色葡萄球菌带菌率可能较高,应引起有关部门的重视。     

Isolation and identification of mycobacteria from cattle slaughtered in Pakistan.

Specimens of lung, liver and mesenteric lymph node from cows and buffaloes slaughtered in the Lahore area were cultured to investigate the type of mycobacteria involved in bovine tuberculosis. Employing the concentration method, 56 out of 530 cattle were found to be culture positive for acid-fast bacteria, 48 being Mycobacterium bovis and eight atypical mycobacteria. No M tuberculosis or M avium was isolated. Most of the isolated M bovis strains were found to be highly virulent for rabbits.     

Impact of sawdust and wood shavings in bedding on pig tuberculous lesions in lymph nodes, and IS1245 RFLP analysis of Mycobacterium avium subsp. hominissuis of serotypes 6 and 8 isolated from pigs and environment

Among 25,027 slaughter pigs raised in two farms, tuberculous lesions were detected in the lymph nodes of 898 (3.6%) of them. Tuberculous lesions were most commonly found in the mesenteric (601; 2.4%) and head (451; 1.8%) lymph nodes. Mycobacteria were isolated from 49 of 120 randomly selected mesenteric, head and bronchial lymph nodes with diagnosed tuberculosis originating from both farms. Forty six Mycobacterium avium subsp. hominissuis, one M. chelonae and two M. fortuitum isolates were found in the lymph nodes of pigs. No statistically significant difference was detected between farms A and B for isolation rates of mycobacteria from the lymph nodes of pigs and their species composition. To investigate the source of the pigs' infections, culture examinations of 117 samples from the external environment were performed. Mycobacteria were isolated from 25 samples from the external environment (21.4%). Mycobacterial isolates were also detected in eleven (91.7%) and two (16.7%) of 12 used sawdust and 12 of non-used (fresh) sawdust samples, respectively. None of 12 wood shavings was culture-positive. Twelve of 13 sawdust isolates were classified as M. a. hominissuis of serotypes 6 and 8 and genotype IS901- and IS1245+; the remaining isolate was classified as species M. fortuitum. Other conditionally pathogenic mycobacteria were only isolated from 12 of the remaining 81 samples from the external environment (excluding bedding). A total of eight isolates (two pig and six sawdust samples originating from farms A and B) were examined by IS1245 restriction fragment length polymorphism (IS1245 RFLP) analysis. These isolates produced five distinct IS1245 RFLP types with more than 20 bands. Based on identical IS1245 RFLP types of one pig isolate and two isolates of used sawdust from farm A, we have concluded that contaminated sawdust was the source of mycobacterial infection for pigs in our study.     

RESPONSE OF CATTLE TO INOCULATION WITH ATYPICAL MYCOBACTERIA ISOLATED FROM SOIL

Nine strains of atypical mycobacteria and a strain of the rhodochrous taxon, originally isolated from soil samples collected on the subcoastal plains of the Northern Territory, were inoculated into cattle. Each strain was injected subcutaneously into one animal and into a mesenteric lymph node of another. At 4 and 10 weeks after inoculation, the cattle were tuberculin tested with bovine PPD tuberculin, avian PPD tuberculin and the appropriate homologous PPD tuberculin. Six strains induced a significant level of sensitivity to bovine PPD at the 4-week test, but only one animal gave a similar response at the 10-week test. In general, the level of sensitivity to all tuberculins declined between the 4-week and 10-week tests. At both tests the response to avian PPD was equal to, or exceeded, that to bovine PPD. The inoculation of each of the 10 strains resulted in the production of tuberculous granulomas at the subcutaneous sites and similar lesions were produced at the mesenteric lymph node site in response to 2 strains. Mycobacteria were re-isolated from 11 cattle and represented 7 strains. The significance of the soil as a reservoir of atypical mycobacteria and other organisms capable of inducing sensitivity to bovine PPD is discussed.     

Cultural and biochemical characteristics of Mycobacterium paratuberculosis isolated from goats in Norway.

In Norway a variant of Mycobacterium paratuberculosis occurs which causes disease in goats but very seldom in sheep and cattle. Cultural and biochemical characteristics of this variant are investigated by comparing different pre-treatment methods and culture media for primary isolation and by subjecting a number of strains to different enzymatic and biochemical tests. Decontamination of materials with 5% oxalic acid and 0.1% benzalkonium chloride and culture on Dubos, Finleyson’s and Herrold’s medium was tested. The investigations showed that the combination oxalic acid decontamination/Dubos’ medium is most suitable for isolation of the goat-pathogenic variant.The morphology of the colonies was also most easily studied after culture on Dubos’ medium from material pre-treated with oxalic acid. The biochemical tests were found to be poorly suitable for the identification of M. paratuberculosis and for its differentiation from other mycobacteria.Mycobactin dependence for growth seems not to be absolute as a few goat strains produced growth on Dubos’ medium without mycobactin. However, growth was in all cases far better in the presence of mycobactin.     

猪肺炎支原体YN200901株的分离鉴定

采集疑似猪肺炎支原体病理变化的肺脏,接种到改良牛心培养基、猪胃消化液培养基进行培养,经瑞特染色、镜检,菌落狄氏染色、PCR鉴定及序列比对、生化试验鉴定,抗猪肺炎支原体血清生长抑制试验,分离获得一株猪肺炎支原体云南地方流行菌株,命名为YN200901。试验结果为云南本地株猪肺炎支原体的后续研究提供了物质基础。     

An investigation of the effects of secondary processing on Mycobacterium spp. in naturally infected game meat and organs

The risk for humans to contract bovine tuberculosis through the consumption of undercooked game meat as well as biltong (traditionally dried game meat) is a concern. The survival potential of Mycobacterium bovis during the cooking and drying processes was researched in a preceding study on beef and the positive results compelled the authors to investigate the results with a similar preliminary study on game meat. Muscular, lymphatic and visceral tissues from skin test positive African buffalo (Syncerus caffer) and greater kudu (Tragelaphus strepsiceros) with tuberculous lesions were collected from the Hluhluwe iMfolozi Park during the park's culling programme. The different tissues were exposed to cooking and the muscular tissue to the drying process prior to culture. All acid-fast isolates were analysed by polymerase chain reaction for the presence of Mycobacterium bovis. All tissues were found negative for Mycobacterium bovis but non-tuberculous mycobacteria were isolated from kidney, liver, heart and lymph nodes. The results showed that these processes will kill Mycobacterium bovis but the unexpected recovery of non-tuberculous mycobacteria suggests possible survival and resistance characteristics of these strains which might be of veterinary public health interest.     

Species identification of non-tuberculous mycobacteria from humans and cattle of Chad

In Chad, during a study on tuberculosis in humans and cattle, 52 non-tuberculous mycobacteria (NTM) strains were isolated. By means of INNO-LiPA, PRA-hsp65 amplification and sequencing of 16S rDNA, NTM species of 25/52 isolates were identified. M. fortuitum complex (8) was the most frequent species, followed by M. nonchromogenicum (4) and M. avium complex (4). PRA method could identify M. fortuitum 3rd variant among isolates derived from cattle specimens. This finding could confirm the existence of farcy in the Chadian cattle population as M. fortuitum 3rd variant and putitative pathogen M. farcinogenes can't be distinguished by the methods used in this study. Half of the NTM isolates could not be specified and we considered them as contaminants from the environment.